Nitrilase mutant with improved catalytic activity and reaction specificity and application

A technology of nitrilase and mutants, applied in the field of bioengineering, can solve the problems of production process and production cost increase, achieve the effects of reducing industrial production costs, improving nitrilase activity and reaction specificity, and good application prospects
CN112359036AActive Publication Date: 2021-02-12ZHEJIANG UNIV OF TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
ZHEJIANG UNIV OF TECH
Publication Date
2021-02-12

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Abstract

The invention discloses a nitrilase mutant and application thereof in hydrolysis of racemic isobutyl butyronitrile to synthesize a pregabalin chiral intermediate (S)-3-cyano-5-methyl hexanoic acid andbelongs to the technical field of bioengineering. The nitrilase mutant BaNITmut / V82L with the amino acid sequence as shown in SEQ ID NO.4 or the double mutant BaNITmut / V82L / C237S with the amino acidsequence as shown in SEQ ID NO.6 can catalyze high-concentration IBSN hydrolysis, and amide byproducts are remarkably reduced. The nitrilase activity and the reaction specificity are improved througha directed evolution technology, industrial production cost is greatly reduced, and the method has a good application prospect in industrial production of the pregabalin chiral intermediate (S)-3-cyano-5-methyl caproic acid.
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Description

technical field

[0001] The invention belongs to the technical field of bioengineering, and relates to a mutant of a plant-derived nitrilase, a coding gene, and a chiral intermediate (S)-3-cyano in the hydrolysis of racemic isobutylsuccinonitrile to synthesize pregabalin - Application of 5-methylhexanoic acid. Background technique

[0002] Pregabalin, whose chemical name is (S)-3-aminomethyl-5-methylhexanoic acid, has the structural formula (I), and is the 3-position of the inhibitory neurotransmitter γ-aminobutyric acid (GABA). Isobutyl substituents (Angew. Chem. Int. Ed., 2008, 47:3500-3504). Pregabalin is the drug of choice for treating conditions such as nerve pain, anxiety, and epilepsy. Therefore, it is of great significance to develop a green, efficient, and highly optically pure pregabalin synthesis process.

[0003]

[0004] (S)-3-cyano-5-methylhexanoic acid (CMHA) is the key chiral intermediate of pregabalin, and this compound can be synthesized into S-form pr...

Claims

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